3alpha-succinoxy-11-hydroxy-12-keto-delta9,(11)-cholenic acid and its derivatives



United States Patent 3ot-SUCCINOXY-lI-HYDROXY-IZ-KETO-A 1 CHOLENIC ACID AND ITS DERIVATIVES Julien Warnant, Neuilly-sur-Seine, Gerard Noe, Noisy -le-Sec, and Genevieve Rousseau, Errghien-les- Bains, France, assignors to Les Laboratoires Francais de Chimiotherapie, Paris, France, a body of France N Drawing. Application February 26, 1957 Serial No. 642,331

Claims priority, application France February 29, 1956 3 Claims. (Cl. 260-3971) The present invention relates to new derivatives of dihydroxy keto cholenic acid and more particularly to 3- succinoxy-ll-hydr0xy-12-keto-A -cholenic acid and its salts, and to a process of making same.

It is one object of the present invention to provide a new derivative of a dihydroxy keto cholenic acid and more particularly to provide 3-succinoxy-l1-hydroxy-12- keto-A -cholenic acid and salts thereof, which compounds have proved to be valuable agents in modifying the lipoprotein equilibrium of the serum.

Another object of the present invention is to provide a simple and eifective process of making said 3-succinoxy- 1l-hydroxy-l2-keto-A -cholenic acid and its salts.

Still another object of the present invention is to provide a therapeutically useful preparation for modifying the lipoprotein equilibrium of the serum which preparation is characterized by a content of 3-succinoxy-11-hydroxy-l2-keto-A -cholenic acid or, respectively, its salts Other objects of the present invention and advantageous features thereof will become apparent as the description proceeds.

It is known that normal serum contains, as an average, between 1.5% and 2.0% of total cholesterol, i.e. of free cholesterol and esterified cholesterol. These figures have the tendency to increase in the course of the evolution of certain nutritional disorders or because of certain diseases such as atherosclerosis. It is, therefore, of great importance, to place at the disposal of the medical profession an agent which is capable of reducing, at least to a certain extent, pathological accumulations and concentrations of cholesterol in the blood.

In our co-pending application Serial No. 591,278, filed June 14, 1956, and entitled New Derivative of a Hydroxy-Diketocholanic Acid and a Method of Preparing Same, compounds have been described which are capable of progressively decreasing the cholesterol content of the blood when injected intravaneously. The preferred compound for said purpose is 3-succinoxy-11,12-diket0cholanic acid.

In principle the present invention deals with the enolic form of said acid, namely with 3-succinoxy-11-hydroxy- 12-keto-A -cholenic acid of Formula 1.

CH CH O 0 CH2 COO 3132 OOH Said acid, especially in the form of its salts with suitable bases, permits, by oral administration, to restore quite rapidly the cholesterol content of the blood of chickens which had previously been increased by the application of a feed rich in oil and cholesterol, to its normal cholesterol content. Such a favorable result is achieved when treating chickens having an increased cholesterol blood level for a few weeks with said acid. The treatment has the further advantage that the values obtained in the Kunkel test are lowered and that the plasma is clarified. The toxicity of 3-succinoxy-1l-hydroxy-12-keto-A cholenic acid is remarkably low.

The new acid is preferably obtained according to the present invention by treating 3u,l1-dihydroxy-12-keto- A -cholenic acid in solution in a suitable solvent with an agent capable of introducing the succinic acid group, such as succinic acid anhydride, in the presence of a condensing agent, such as a tertiary base, for instance, pyridine, and isolating the resulting 3a-succinoXy-ll-hydroxy- 12-keto-A -cholenic acid.

The 3oz succinoxy ll-hydroxy-12-keto-A -cholenic acid is preferably used in the form of its salts which are highly soluble in water such as salts with an inorganic or organic base. The free acid or its salts with bases are administered in the form of capsules, pellets, tablets, dragees, or other preparations permitting oral administration.

A preferred method of introducing the succinyl group into the 3u,ll-dihydroxy-12-keto-A -cholenic acid consists in dissolving said acid in pyridine which serves as solvent as well as a condensing agent, adding to said solution succinic acid anhydride, heating the mixture for one to two hours at a temperature of about C., and isolating 3a-succinoxy-ll-hydroxy-l2-ketoA -cholenic acid by the conventional methods of Washing, extraction, and crystallization.

The salts of the resulting 3a-succinoxy-11-hydroxy-l2- keto-A -cholenic acid are prepared by neutralizing a solution of said acid in a suitable solvent by means of a selected inorganic or organic base. The resulting salt is separated by centrifuging, filtration, or evaporation to dryness of the reaction medium.

30 11 dihydroxy l2-keto A -cholenic acid, which serves as starting material for carrying out the process according to the present invention, is obtained according to Wintersteiner and Moore, Journal Biological Chemistry, volume 162, page 731 (1946).

The following examples serve to illustrate the present invention and especially the process of making said acid and its salts without, however, limiting the same thereto. It is, of course, possible to vary the solvents, reactants, reaction conditions, temperature and duration, and the like without departing from the principles of the present invention as they are set forth herein and in the claims annexed hereto. It is also possible to prepare 3oc,11-dihydroxy-l2keto-A -cholenic acid by any other process than that of the above cited reference, for instance, by alkaline hydrolysis of a derivative of 3a-hydroxy-l1,12- diketocholanic acid, such as 3a-acetoxy-ll,l2-diketo cholanic acid methyl ester, 3ot-formyloxy-l1,12-diketo cholanic acid, or 3u-succinoxy-11,l2-diketo cholanic acid.

EXAMPLE 1 Succinylation of 3a,]1-dihydroxy-12-keto-A -cholenic acid -15 g. of 30,11-dihydroxy-12-keto-A -cholenic acid and 4.1 g. of succinic acid anhydride are dissolved in 15 cc. of anhydrous pyridine. The solution is stirred at a temperature of 95 C. for one hour. The resulting reaction mixture is cooled to 30 C. and is then poured, while stirring, into a mixture of 225 cc. of ice-water and 25 cc.

of concentrated hydrochloric acid. The mixture is allowed to stand overnight at room temperature, filtered, washed with water, dried, and re-crystallized from 3 parts by volume of 50% acetic acid, yielding thereby 3oc-SUC- oinoxy-l l-hydroxy-l2-keto-A -cholenic acid with a yield of 72% of the theoretical yield. Melting point: 240 C.; rotary power -[aJ =+87:2 (concentration: 0.5% in acetone). The acid is obtained in the form of colorless crystals which are insoluble in ether, benzene, and chloroform and slightly soluble in water, acetone, and cold ethanol. Its alcoholic solution yields an intense green color with ferric chloride due to its enolic structure. It exhibits an ultraviolet absorption maximum at 281 mg characteristic for B-unsaturated a-hydroxy-ketones.

EXAMPLE 2 Di-sodium salt of 3a-succin0xy-11-hydr0xy- J2-ket0-A -chlenic acid 5 g. of 3a-succ'in'oxy-ll-hydroxy-12-keto-A -cholenic acid are suspended at 65 C. in 10 cc. of water. The suspension is stirred, carbon dioxide is bubbled therethrough, and 33 cc. of an aqueous solution of sodium bicarbonate is added in small portions thereto. The resulting solution is evaporated to dryness in a vacuum at a temperature of 4550 C.; 5.4 g. of-the disodium salt of 3a-succinoxy- 1l-hydroxy-12-keto-A -cholenic acid are obtained. The salt forms a white powder, soluble to about 25% in water at 20 C. and having a rotatory power [a] =-|-90i2 (concentration: 1% in water).

When using, in place of-sodium bicarbonate equimolecular amounts of potassium bicarbonate, and proceeding otherwise in the same manner 'as described in Example 2, the corresponding potassium salts are obtained.

Especially suitable salts with organic bases are salts obtained by reaction with alkanolamines such as ethanolamine, diethanolamine, triethanola'min'e, ethylene diamine.

As stated above, the new acid and its salts are orally effective therapeutic agents for reducing a high cholesterol blood level and normalizing 'same.

The compounds are preferably administered in the form of tablets, pellets, pills, dragees, powders, solutions, emulsions, suspensions, or other orally administrable preparations. The new compounds are preferably used in their dilute form, thus, allowing better and more economical use to be made thereof.

In the administration of such compounds in capsules as powders, a fine uniform dispersion of the active product throughout said powder is desirable. Such a fine dispersion can be achieved, for instance, by intimately mixing 4 and milling the compound in a ball mill with a solid, pulverulent extending agent to the desired degree of fineness, or by impregnating the already milled, finely powdered solid carrier with a mixture of the active compound in water or other suitable solvents and then removing the water or solvent.

When preparing tablets, pills, dragees, and the like shaped solid preparations for oral administration, the commonly used diluting agents, binders, lubricants and the like tableting adjuvants are employed, such as sugar, lactose, talc, starch, bolus alba, as binders pectin, gelatin, gum arabic, methyl cellulose, yeast extract, agar, tragacanth, and as lubricants stearic acids, magnesium stearate, talc, and others.

The content of active compound in such preparations may vary. It is, of course, of advantage, that the active compound be present in said preparations in such an amount that a suitable dosage will be ensured. The unit dose should contain not less than 10% of the active compound. The preferred amounts tobe employed in tablets and the like shaped solid preparations are between about 10% and about 50% of the weight of the unit. To use greater amounts, is, of course, also possible, although administration of suitable doses becomes somewhat cumbersome. The maximum single dose is about 500 mg. and the maximum daily dose is about 1500 mg. The preferred single dose is between about 100 mg. and about 300 mg. and the preferred daily dose is between about 200 mg. and about 1000 mg. Administration of the new acid and its salts is continued until the cholesterol blood level is substantially normal.

It is, of course, also possible to add the acid and its salts to food, especially to the fats used in preparing meals, or

- to flour used in preparing bread and other baked goods.

We claim:

1. A compound selected from the group consisting of 3a-succinoxy-l1-hydroxy-12-keto-A -cholenic acid and its alkali metal salts and salts with' lower alkanol amines and ethylene diamine, said salts being tolerated by the human system.

2. 3 a-succinoxy-l l-hydroxy-l2-keto-A -cholenic acid.

3. The disodium salt of 3a-succinoxy-1l-hydroxy-lZ- keto-rl -cholenic acid.

References Cited in'the file of this patent UNITED STATES PATENTS 2,745,829 Archer May 15, 1956 

1. A COMPOUND SELECTED FROM THE GROUP CONSISTING OF 3A-SUCCINOXY-11-HYDROXY-12-KETO-$9,11-CHOLENIC ACID AND ITS ALKALI METAL SALTS AND SALTS WITH LOWER ALKANOL AMINES AND ETHYLENE DIAMINE, SAID SALTS BEING TOLERATED BY THE HUMAN SYSTEM. 